Devices and Methods for Particle Solution Testing

ABSTRACT

A method for withdrawing a solution comprising a plurality of particles from a vial is disclosed. The method includes agitating the vial at a first predetermined mixing speed to suspend the plurality of particles in the solution. The method also includes withdrawing, during the agitating, a first volume of the solution from the vial with a pipette, such that an end of a pipette tip is positioned at a first predetermined depth within the vial.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent Application No. 63/288,397, filed on Dec. 10, 2021, U.S. Provisional Patent Application No. 63/288,408, filed on Dec. 10, 2021, U.S. Provisional Patent Application No. 63/288,386, filed on Dec. 10, 2021, and U.S. Provisional Patent Application No. 63/288,378, filed on Dec. 10, 2021, each of which is hereby incorporated by reference in its entirety. Additionally, this application hereby incorporates by reference the following U.S. Patent Applications, each filed on the same date as the present application: U.S. patent application Ser. No. ______, titled “Devices and Methods for Pipette Alignment” (identified by Attorney Docket No. 21-0308-US), U.S. patent application Ser. No. ______, titled “Devices and Methods for Particle Solution Preparation” (identified by Attorney Docket No. 21-1234-US), and U.S. Patent Application No. titled “Devices and Methods for Particle Mixing” (identified by Attorney Docket No. 21-1582-US).

FIELD OF THE DISCLOSURE

The present disclosure involves systems and methods for withdrawing a solution containing particles from a vial utilizing a shaker plate system. Namely, devices and methods of the disclosure stabilize vials on a shaker plate and position an end of a pipette tip at a series of predetermined depths during agitation and withdrawing of the solution.

BACKGROUND

Assays (including immunoassays) can be conducted utilizing a variety of different solutions, including solutions containing particles to assist in performing the assays.

SUMMARY

In some examples, particles such as paramagnetic beads, polystyrene particles, or the like, can be suspended within a solution that can be withdrawn with a pipette for testing and identification of components in a sample. In some configurations, different particles may be configured to detect different antibodies, antigens, proteins, or the like. By detecting different, antibodies, antigens, proteins, or the like, the particles can be utilized to perform multiple simultaneous assays on a single sample. To increase the accuracy of assay test results, it is desirable to withdraw approximately the same number of particles during each withdrawing event and, prior to withdrawing, ensure that the particles are dispersed evenly throughout the solution.

When operators manually withdraw solution using a pipette, the results of the assay may be inconsistent. For example, an operator's positioning of the pipette within a vial of the solution can significantly affect the type, number, and consistency of particles withdrawn from the solution. Further, any inconsistency of the pipette from one withdrawing event to another can also impact the accuracy and precision of the assay results. For example, in some configurations different particles within the solution are utilized to perform different assays. As the homogeneity of the particles within the solution decreases, the likelihood of withdrawing an uneven distribution of the different particles increases, such that particles associated with a specific assay may be over or under represented in a volume of solution withdrawn, which can impact the result of the specific assay. Accordingly, manual withdrawals of the solution are subject to variability between withdrawing events and/or operators and, thus, can degrade the accuracy and precision of associated assay results.

In an example, a method is described for withdrawing a solution comprising a plurality of particles from a vial. The method comprises agitating the vial at a first predetermined mixing speed to suspend the plurality of particles in the solution. The method also comprises withdrawing, during the agitating, a first volume of the solution from the vial with a pipette such that an end of a pipette tip is positioned at a first predetermined depth within the vial.

In another example, a non-transitory computer-readable medium is described, having stored thereon program instructions that, upon execution by a controller cause a controller to perform a set of operations. The set of operations comprises agitating a vial at a first predetermined mixing speed to suspend a plurality of particles in a solution. The set of operations also comprises withdrawing, during the agitating, a first volume of the solution from the vial with a pipette, such that an end of a pipette tip is positioned at a first predetermined depth within the vial.

The features, functions, and advantages that have been discussed can be achieved independently in various examples or may be combined in yet other examples. Further details of the examples can be seen with reference to the following description and drawings.

BRIEF DESCRIPTION OF THE FIGURES

The above, as well as additional features will be better understood through the following illustrative and non-limiting detailed description of example embodiments, with reference to the appended drawings.

FIG. 1 illustrates a simplified block diagram of an example computing device, according to an example embodiment.

FIG. 2A illustrates a vial rack, pipettes, and a shaker plate, according to an example embodiment.

FIG. 2B illustrates the vial rack, pipettes, and the shaker plate of FIG. 2A, according to an example embodiment.

FIG. 2C illustrates the vial rack and the shaker plate of FIG. 2A, according to an example embodiment.

FIG. 2D illustrates a top view of the vial rack of FIG. 2A, according to an example embodiment.

FIG. 2E illustrates a perspective view of the vial rack of FIG. 2A, according to an example embodiment.

FIG. 2F illustrates a side view of the vial rack of FIG. 2A, according to an example embodiment.

FIG. 2G illustrates another side view of the vial rack of FIG. 2A, according to an example embodiment.

FIG. 3A illustrates a vial and an end of a pipette tip of the pipette of FIG. 2A at a first predetermined depth, according to an example embodiment.

FIG. 3B illustrates the vial and the end of the pipette tip of the pipette of FIG. 3A at a second predetermined depth, according to an example embodiment.

FIG. 3C illustrates another vial and an end of a pipette tip of the pipette of FIG. 2A at a first predetermined depth, according to an example embodiment.

FIG. 3D illustrates the vial and the end of the pipette tip of the pipette of FIG. 3C at a second predetermined depth, according to an example embodiment.

FIG. 4A illustrates a sample of prepared solution containing particles according to an example embodiment.

FIG. 4B illustrates an image of the sample of prepared solution containing particles of FIG. 4A, according to an example embodiment.

FIG. 4C illustrates a composite image of the sample of solution containing particles of FIGS. 4A and 4B and an associated graphical user interface, according to an example embodiment.

FIG. 4D illustrates an annotated version of the composite image of FIG. 4C and an associated graphical user interface, according to an example embodiment.

FIG. 5A illustrates particle count assay results at varying shaker plate speeds.

FIG. 5B illustrates particle count assay results at a mixing speed of 1000 revolutions per minute.

FIG. 5C illustrates particle count dispersion without a buffer read phase.

FIG. 5D illustrates particle count dispersion after conducting a buffer read phase.

FIG. 5E illustrates particle count readings using an automated counting program compared to a manual counting protocol after a solution is prepared and an associated graphical user interface according to an example embodiment.

FIG. 6 illustrates a method, according to an example embodiment.

All the figures are schematic, not necessarily to scale, and generally only show parts which are necessary to elucidate example embodiments, wherein other parts may be omitted or merely suggested.

DETAILED DESCRIPTION

Example embodiments will now be described more fully hereinafter with reference to the accompanying drawings. That which is encompassed by the claims may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided by way of example. Furthermore, like numbers refer to the same or similar elements or components throughout.

Within examples, the disclosure is directed to devices and methods for withdrawing samples of a solution containing one or types of particles utilizing a shaker plate system. In particular, as described in the example embodiments, the one or more types of particles may include one or more of the following: microbeads, microparticles, micropellets, microwafers, paramagnetic beads, microparticles, paramagnetic microparticles, or the like. In embodiments, each of the particles may include an identifying feature, such as a bar code, a nickel bar code, and/or identifying features other than bar codes, including a color, a shape, an alphanumeric symbol, and/or the like. Moreover, the particles may be magnetic or paramagnetic. Particles suitable for use in the disclosure are capable of attachment to other substances such as derivatives, linker molecules, proteins, nucleic acids, or combinations thereof. The capability of the particles to be attached to other substances can result from the particle material as well as from any further surface modifications or functionalization of the particle. The particles can be functionalized or be capable of becoming functionalized in order to covalently or non-covalently attach proteins, nucleic acids, linker molecules or derivatives as described herein.

For example, the surface of the particle can be modified or functionalized with amine, biotin, streptavidin, avidin, protein A, sulfhydryl, hydroxyl and carboxyl. Particles may be spherical or other shapes, may be light transmissive and may be digitally coded such as for example, by an image that provides for high contrast and high signal-to-noise optical detection to facilitate identification of the bead. To the extent an image is present, the image may be implemented by a physical structure having a pattern that is partially substantially transmissive (e.g., transparent, translucent, and/or pervious to light), and partially substantially opaque (e.g., reflective and/or absorptive to light) to light. The pattern of transmitted light is determined (e.g., by scanning or imaging), and the code represented by the image on the coded bead can be decoded. Various code patterns, such as circular, square, or other geometrical shapes, can be designed as long as they can be recognized by an optical decoder. Examples of these one or more types of particles may be found at: U.S. Pat. Nos. 7,745,091, 8,148,139, and 8,614,852.

Additionally or alternatively, the one or more types of particles may comprise one or more materials, including one or more of the following: glass, polymers, polystyrene, latex, elemental metals, ceramics, metal composites, metal alloys, silicon, or of other support materials such as agarose, ceramics, glass, quartz, polyacrylamides, polymethyl methacrylates, carboxylate modified latex, melamine, and Sepharose, and/or one or more hybrids thereof. In particular, useful commercially available materials include carboxylate modified latex, cyanogen bromide activated Sepharose beads, fused silica particles, isothiocyanate glass, polystyrene, and carboxylate monodisperse microspheres. Furthermore, the one or more types of particles may also comprise one or more specific shapes, dimensions, and/or configurations and may be modified for one or more specific uses. For example, the particles may be a variety of sizes from about 0.1 microns to about 100 microns, for example about 0.1, 0.5, 1.0, 5, 10, 20, 30, 40 50, 60, 70, 80 90 or 100 microns. For example, the one or more types of particles may be surface modified and/or functionalized with biomolecules for use in biochemical analysis.

The particles of the disclosure may be used in various homogenous, sandwich, competitive, or non-competitive assay formats to generate a signal that is related to the presence or amount of an analyte in a test sample. The term “analyte,” as used herein, generally refers to the substance, or set of substances in a sample that are detected and/or measured, either directly or indirectly. In various aspects the assays of the disclosure, examples include sandwich immunoassays that capture an analyte in a sample between a binding member (e.g., antibody) attached to the particle and a second binding member for the analyte that is associated with a label. In another example embodiment, the binding member on the particle may be an antigen (e.g., protein) that binds an antibody of interest in a patient sample in order to capture the antibody on the particle. The presence of the antibody can then be detected with a label conjugated to a second binding member specific for an antibody. The second binding member attached to the label may be the antigen conjugated to the label or the binding member may itself be an antibody (e.g., anti-species antibody) that is conjugated the label. In example embodiments, these characteristics may be referred to herein as a “unique identifying feature” of the particle. Other examples are possible. For example, the particle may also bind to a fluorescent tag or label, which may present a “unique identifying feature” of the particle to which the fluorescent tag or label might bind under a fluorescent lighting.

In another example embodiment, the assay methods of the disclosure are competitive immunoassays for detection of antibody in the sample. A competitive immunoassay may be carried out in the following illustrative manner. A sample, from an animal's body fluid, potentially containing an antibody of interest that is specific for an antigen, is contacted with the antigen attached to the particle and with the anti-antigen antibody conjugated to a detectable label. The antibody of interest, present in the sample, competes with the antibody conjugated to a detectable label for binding with the antigen attached to the particles. The amount of the label associated with the particles can then be determined after separating unbound antibody and the label. The signal obtained is inversely related to the amount of antibody of interest present in the sample.

In an alternative example embodiment of a competitive a sample, an animal's body fluid, potentially containing an analyte, is contacted with the analyte conjugated to a detectable label and with an anti-analyte antibody attached to the particle. The antigen in the sample competes with analyte conjugated to the label for binding to the antibody attached the particle. The amount of the label associated with the particles can then be determined after separating unbound antigen and label. The signal obtained is inversely related to the amount of analyte present in the sample.

Antibodies, antigens, and other binding members may be attached to the particle or to the label directly via covalent binding with or without a linker or may be attached through a separate pair of binding members as is well known (e.g., biotin:streptavidin, digoxigenin:anti-digoxiginen).

Assays using these solutions are often conducted over a series of agitation events and withdrawing events. In practice, the particles in the solution may bind together (often referred to as “clumping”) or bind and/or settle on the bottom or sides of a vial. This binding may result in an inconsistent dispersion of the particles in the solution. For example, the relative density of the particles may be lower than the density of the solution, which may result in the particles floating to the top of the solution and inconsistent particle dispersion throughout the solution.

When these particles clump together, they may not be accurately identified or accounted for in the assay. In another example, in instances where the particles bind/settle on the bottom or sides of the vial, the particles may remain in the vial as solution is pipetted out of the vial. To help address this issue, a shaker plate system, which typically includes vials coupled to a shaker plate and pipettes controlled via a programmable controller, can agitate the solution (for example by way of aspiration and/or shaking the vials via the shaker plate) before a withdrawing event in order to more consistently disperse the particles in the solution. However, the mixing speed and mixing pattern of the shaker plate, the stabilization of vials, and the depth of the pipette within the vial during an agitation and/or withdrawing event may affect the consistency of the assay results and/or the type and consistency of particles dispersed within the solution.

To help address these issues, a program for controlling the shaker plate system can be utilized to cause the shaker plate and pipette to perform a preset series of agitation and withdrawing events to improve accuracy and consistency of assay results across both vials and tests, thereby increasing the accuracy of the assay results as compared to assays performed with inconsistent types or numbers of particles. Namely, the controller of the shaker plate system can cause the shaker plate and pipettes to execute a series of agitation and withdrawing events where the mixing speed of the shaker plate is set to a predetermined rotational velocity, pattern (e.g., orbital, linear, etc.), and/or other parameters. For example, the controller of the shaker plate may cause the shaker plate systems to move in a pattern and/or specific tolerance to ensure that one or more components operate within a specific range of parameters in relation to one another. For example, the controller may cause the shaker plate system to position one or more vials such that a pipette is in the horizontal center of the vials and shake the vials in an orbital pattern in an orbit within a predetermined tolerance of the pipette in relation to the vials from the pipette to the walls of the vial in which the pipette is inserted) (e.g., an orbital patter of up to 3 mm).

In example embodiments, certain mixing speeds and patterns may more consistently agitate the solution, which can lead to consistent dispersion of the particles within the solution, thereby producing more consistent particle counts than other mixing speeds and patterns.

Additionally, the controller can cause the pipettes to agitate and withdraw solution at a predetermined depth within the vial and with respect to the depth of the solution within the vial. In example embodiments, the controller may cause the pipettes to agitate and withdraw solution at a predetermined depth within the vial and with respect to the depth of the solution within the vial by controlling one or more components of the systems and devices described here (e.g., the pipettes, the shaker plate system, etc.) and/or one subparts thereof (e.g., one or more motors, actuators, and/or other mechanical components of the pipettes, the shaker plate system, etc.). By doing so, the accuracy of particle count and/or type of particles withdrawn from solution may be improved as compared to systems that do not cause the pipettes to agitate and withdraw solution at a predetermined depth. For example, positioning the end of the pipette tip at or near the center or middle of the volume (e.g., the center line of the height of the solution in the vial or the “vertical center”) may allow the pipette to more effectively agitate the solution to produce more consistent particle counts compared to other positions such as the bottom of the solution or the top of the solution. As such, in example embodiments, the controller causes the pipettes to be positioned such that, during a withdrawing event, the end of the pipette tip is at or near the vertical and/or horizontal center of the volume of the solution or another position that is consistent, on a relative volume basis, for withdrawn from the vial.

Similarly, positioning the end of the pipette tip at or near the center or middle of the horizontal cross section of the vial (the “horizontal center”) may allow the pipette to more consistently aspirate and/or agitate the solution. As used in this disclosure, horizontal indicates a direction transverse to the vertical direction, and extending outward from a vertical centerline of the referenced body (e.g., the horizontal center of a vial stabilized in one position in a vial receptacle). This specific positioning can produce more consistent particle counts compared to other positions, such as the side of the vial. As such, in example embodiments, the pipette alignment device and methods described herein position a pipette such that, during a withdrawing event, the end of the pipette tip is at near the horizontal center of the vial for withdrawal from the vial.

In practice, removal of the contents of a vial may be performed over a series of withdrawing events. For example, a first volume of the solution may be withdrawn, and a first assay conducted. Then a second volume of the solution may be withdrawn, and second assay conducted. This process may be repeated a number of times (e.g., 12). As noted above, during each of the withdrawing events it may be beneficial for the withdrawing tip of the pipette to be in the vertical and/or horizontal center of the solution. However, as the volume of solution is reduced with each withdrawing event, the vertical center of the solution becomes lower within the vial. Thus, attaining consistent vertical positioning of the pipette within the solution as the volume decreases may also provide more consistent assay results (e.g., by improving the consistency of the amount and/or type of particles withdrawn at each withdrawing event over the series of withdrawing event). Further, these agitation and withdrawing events may be performed across multiple vials of solution at once (e.g., eight vials).

Devices and methods of the present disclosure also involve positioning a pipette such that the end of the pipette tip is at or near the vertical and/or horizontal center of the solution or another consistent location (e.g., 30%, 40%, 50%, 60% or 70% of the height of the solution) during each withdrawing event in a series of withdrawing events. While the vertical center of the vial is typically appropriate for most series of withdrawing events, another position within the solution for each withdrawing event may also be appropriate depending on the amount of solution and the type of particles within the vial.

Indeed, in some embodiments, the position of the pipette tip may be adjusted with each withdrawing event in order to accommodate the amounts of solutions and the particles. For instance, as the solution depth diminishes, the vertical center may be too close to the bottom of the vial in order to provide a withdrawal of solution with number of particles that is consistent with the particles from a previous withdrawing event. For example, as the vertical center becomes too close to the bottom of the vial, the particles may cling to the bottom of the vial and/or settle at the bottom of the vial due to one or more factors (e.g., gravity), the withdrawing position may be adjusted to compensate and provide a more homogeneous solution withdrawing evert. For example, once the vertical center becomes too close to the bottom of the vial, the withdrawing position may be adjusted to withdraw at a distance that is great than the vertical center of the remaining solution (e.g. to withdraw a solution that is less concentrated with particles as compared to the solution at the vertical center).

In another example, in vials that contain solution that has not been agitated and/or had any solution withdrawn, the vertical center may be too far from particles settled on the bottom of the vial to adequately mix the solution and particles to provide a homogenous solution. Therefore, while the device and methods of the disclosure are described herein as aligning the pipette tip at the vertical center of the vial, it should be understood that other locations are possible in the series of withdrawing event, including positions that change with each event relative to the amount of solution in the vial (e.g., at 30%, 40%, 50%, 60% or 70% of the height of the solution).

In some embodiments, a vial rack configured to detachably associate with the shaker plate can be utilized to secure and stabilize the vials. Additionally, the vial rack can be utilized to align and/or secure the vials containing the solution with the pipettes of the shaker plate system for withdrawal and/or assay testing. In particular, the vial rack secures and stabilizes the vials during agitation and withdrawing events, so that solution and particle agitation and/or position are consistent during the withdrawing events (e.g., from assay to assay and/or operator to operator) using the same vials over multiple assays and/or using new samples in different vials.

Referring now to the figures, FIG. 1 is a simplified block diagram of an example computing device 100 of a system (e.g., those illustrated in FIGS. 2A-2B, described in further detail below). Computing device 100 can perform various acts and/or functions, such as those described in this disclosure. Computing device 100 can include various components, such as processor 102, data storage unit 104, communication interface 106, and/or user interface 108. These components can be connected to each other (or to another device, system, or other entity) via connection mechanism 110.

Processor 102 can include a general-purpose processor (e.g., a microprocessor) and/or a special-purpose processor (e.g., a digital signal processor (DSP)).

Data storage unit 104 can include one or more volatile, non-volatile, removable, and/or non-removable storage components, such as magnetic, optical, or flash storage, and/or can be integrated in whole or in part with processor 102. Further, data storage unit 104 can take the form of a non-transitory computer-readable storage medium, having stored thereon program instructions (e.g., compiled or non-compiled program logic and/or machine code) that, when executed by processor 102, cause computing device 100 to perform one or more acts and/or functions, such as those described in this disclosure. As such, computing device 100 can be configured to perform one or more acts and/or functions, such as those described in this disclosure. Such program instructions can define and/or be part of a discrete software application. In some instances, computing device 100 can execute program instructions in response to receiving an input, such as from communication interface 106 and/or user interface 108. Data storage unit 104 can also store other types of data, such as those types described in this disclosure.

Communication interface 106 can allow computing device 100 to connect to and/or communicate with another other entity according to one or more protocols. In one example, communication interface 106 can be a wired interface, such as an Ethernet interface or a high-definition serial-digital-interface (HD-SDI). In another example, communication interface 106 can be a wireless interface, such as a cellular or WI FI interface. In this disclosure, a connection can be a direct connection or an indirect connection, the latter being a connection that passes through and/or traverses one or more entities, such as such as a router, switcher, or other network device. Likewise, in this disclosure, a transmission can be a direct transmission or an indirect transmission.

User interface 108 can facilitate interaction between computing device 100 and a user of computing device 100, if applicable. As such, user interface 108 can include input components such as a keyboard, a keypad, a mouse, a touch sensitive panel, a microphone, a camera, and/or a movement sensor, all of which can be used to obtain data indicative of an environment of computing device 100, and/or output components such as a display device (which, for example, can be combined with a touch sensitive panel), a sound speaker, and/or a haptic feedback system. More generally, user interface 108 can include hardware and/or software components that facilitate interaction between computing device 100 and the user of the computing device 100.

Computing device 100 can take various forms, such as a workstation terminal, a desktop computer, a laptop, a tablet, a mobile phone, or a controller.

Now referring to FIGS. 2A-2B, a vial rack 200, a shaker plate 206, one or more vials 208, and one or more pipettes 210 are depicted, according to an example embodiment. FIG. 2C illustrates the embodiments in FIGS. 2A-2B, from an alternative angle and without the explicit depiction of pipettes 210. The vial rack 200 includes a base 204 coupled to one or more vial receptacles 202, suitable for holding vials 208.

In examples, each of the vial receptacles 202 are configured to hold one or more vials 208 (e.g., one vial 208 for every vial receptacle 202). In some examples, the vial receptacles 202 are configured to hold a variety of vials, some or all of which may meet Society for Biomolecular Screening (“SBS”) standards or other industry standard for laboratory equipment. In the embodiment shown in FIGS. 2A-2C, the vial receptacles 202 are aligned to position the vials 208 such that an SBS standard pipettes (for example, pipettes 210) can be inserted and take withdrawals from each of the vials. Namely, the vial receptacles 202 are designed to space vials similarly to standard SBS pipette tips, such as the Hamilton® SPAN8-channel pipetting mechanism. Additionally, although various components of the systems and/or methods described herein may refer to SBS-compliment components, they are described for the purposes of illustrating example embodiments and other, additional components may be utilized.

For example, although the vial receptacles 202 are illustrated in FIGS. 2A-2C as sharing interlocking walls in one direction, vial receptacles 202 may be implemented in one or more additional or alternative configurations. For example, vial receptacles 202 may share interlocking walls in one or more other directions (e.g., in the space between the four vial receptacle rows) to improve structural strength between the vial receptacles in each row and/or may be implemented in various proportions compared to the embodiments illustrated in FIGS. 2A-2C. For example, the vial receptacles 202 may be taller or shorter than the embodiments illustrated in FIGS. 2A-2C (e.g., to improve the rigidity and/or grip of the vial receptacle on the vial inserted into each vial receptacle). In some embodiments the vial receptacles 202 may incorporate one or more additional materials to provide additional functionality. For example, vial receptacles 202 may be lined with a rubber and/or foam that might allow the vials to more securely seat in the vial receptacles 202 as compared to vial receptacles that do not include a liner. Additionally, these materials lining the vial receptacles 202 may absorb one or more mechanical actions (e.g., shaking) of the pipette or other components, thereby reducing transfer of mechanical energy to the vials during withdrawing events. In some embodiments, one or more vials may be molded into the vial receptacles 202.

In some examples, the vial receptacles 202 may be adjustable to hold different sizes, numbers, or types of vials. For example, although the vial receptacles 202 are shown in FIG. 2B as semi-circular shape (e.g., to surround standard SBS vials) with shared walls relative to each other in one direction, these vial receptacles may take a different shape (e.g., oval, square) and/or share walls in one or more other configurations (e.g., share a wall with other vial receptacles in a different direction). For example, each individual vial receptacle within the vial receptacles 202 may not share a wall with any other vial receptacle (i.e., each vial receptacle's wall may be spaced apart from the vial receptacles' walls) or each vial receptacle may share walls with every vial receptacle that surrounds the individual vial receptacle.

Additionally or alternatively, the height of the vial rack 200 (or individual components thereof) may be adjustable to accommodate different types of pipettes and/or pipette tips. In some examples, the height of the vial rack 200 may also be adjusted to position the end of the pipette tip at one or more predetermined depths during a withdrawing event. In example embodiments, the height of the vial receptacles 202 may be adjustable to accommodate different types of pipettes and/or pipette tips and/or further stabilize the vial during withdrawing events (e.g., increasing the wall height of the vial receptacle may further secure the vial placed therein).

The vial rack 200 may also be detachably associated with the shaker plate 206 by way of a fastening mechanism, such as a screw or other threaded connection in a manner that is not destructive to the vial rack 200, the vial receptacles 202, the shaker plate 206, the pipettes 210, and/or other components of the illustrated devices. In some examples, the dimensions of the surface of the vial rack 200 are the same or substantially similar with the shaker plate 206.

In examples, the shaker plate 206 and pipettes 210 are part of a shaker plate system which includes a computing device, such as computing device 100. As described above, a computing device 100 can be implemented as a controller, and a user of the controller can use the controller to control a shaker plate 206 and pipettes 210. The shaker plate 206 and pipettes 210 communicably coupled with a controller, such as computing device 100, and may communicate with the controller by way of a wired connection, a wireless connection, or a combination thereof.

In examples, the controller can execute a program that causes the shaker plate 206 and pipettes 210 to perform a series of withdrawing events by way of a non-transitory computer-readable medium having stored program instructions. These program instructions include, for example, shaker plate 206 mixing speed (e.g., rotational velocity), mixing pattern, and pipette 210 depth with respect to the vials 208 and/or the solution within the vials 208. Certain mixing speeds, mixing patterns, and pipette placement may more effectively agitate the solution to produce more accurate and consistent particle counts than other mixing speeds, patterns, and pipette tip positions.

For example, the controller program instructions can include shaking the shaker plate 206 at a first predetermined speed while the vial rack 200 is coupled to the shaker plate 206 and includes at least one vial containing a solution in a vial receptacle 202. This first predetermined speed corresponds to a first agitating and withdrawing event of the solution in the vial. Further, in practice, shaking the shaker plate 206 at this first predetermined speed suspends a plurality of particles (e.g., paramagnetic beads and/or polystyrene particles) within the solution.

In some examples, the controller can cause the shaker plate 206 to shake in orbital pattern, consequently agitating the vials 208 and solution in the orbital pattern. In other examples, the controller can cause the shaker plate 206 to shake in a linear pattern, consequently agitating the vials 208 and solution in the linear pattern.

Further, in some examples, the program instructions can include shaking the shaker plate 206 at various mixing speeds, over a variety of mixing patterns (e.g., orbital mixing patterns, linear mixing patterns, etc.). Furthermore, the program instructions can include shaking the shaker plate 206 and agitating the vials 208 and solution within a predetermined range of mixing speeds. For example, in one embodiment, the program instructions may include shaking the shaker plate 206 at a mixing speed between about 900 revolutions per minute (rpm) and about 1100 rpm. As described in more detail with respect to FIG. 6A, this mixing speed range is desirable to suspend the particles without forming bubbles in the solutions, which could negatively impact the assay. Regarding these embodiments, one or more operators may attach the vial rack 200 to shaker plate 206 and then set a mixing speed and/or mixing pattern on the shaker plate 206 prior to the pipettes 210 being inserted in vial 208.

In some embodiments, the controller causes the pipettes 210 to be positioned such that the corresponding pipette tip is at a first predetermined depth within the corresponding vial 208 during the agitation and withdrawing. This predetermined depth may correspond to the volume of solution within the vial 208. For example, the predetermined depth may be at or near the vertical center of the solution which may more effectively agitate the solution to produce more accurate and consistent particle counts for the particles in the solution (e.g., paramagnetic bead counts).

As these withdrawing events are performed in a sequence, in some examples, the pipette is configured to withdraw a predetermined volume of the solution from within the vial at the first predetermined depth. Without being bound by theory, reducing the volume of the solution reduces the height of the top of the solution within the vial, and understanding the geometry of the vial, withdrawing a predetermined volume of the solution will lower the top of the solution to a first predetermined height in the vial. Knowing this predetermined height and/or withdrawing a predetermined volume may be beneficial for aligning the end of the pipette tip to at or near the vertical and/or horizontal center in subsequent withdrawing events.

In this way, the predetermined depths correspond to the parameters of a series of withdrawing events, such as the volume of solution within the vial during a withdrawing event. In particular, one or more predetermined depths of the end of the pipette tip in the solution correspond to the one or more volumes of solution withdrawn over a series of withdrawing events. As described above it is often desirable to position the end of the pipette tip at or near the vertical center of the solution, as well as position the end of the pipette tip at or near the horizontal center of the solution. For example, the volume of solution will decrease after each withdrawing event and one or more predetermined depths correspond to the anticipated vertical center of the solution after a predetermined volume of solution is removed from the vial after each withdrawing event.

In embodiments, the anticipated vertical center of the solution may correspond to the anticipated vertical center of the solution at the beginning of the withdrawing event. In other examples, the anticipated vertical center of the solution may correspond to the anticipated vertical center of the solution at the conclusion of the withdrawing event. In other examples, the anticipated vertical center of the solution may correspond to the anticipated vertical center of the solution at the conclusion of a particular withdrawing event in a series of withdrawing events (e.g., the vertical center of the solution after a first withdrawing event in a series of two withdrawing events). In any event, positioning the end of the withdrawing tip of the pipette at or near the center of the solution positions the pipette tip to provide consistent agitation of the solution, including any particles therein, which is desirable for testing.

In embodiments, the controller may cause the pipettes 210, during the agitating to withdraw a predetermined volume of solution. This predetermined volume of solution determines the pipette 210 position for subsequent agitation and withdrawing events.

This sequence of agitation and withdrawing of the solution may be repeated a number of times. For instance, in some examples, during a second agitating event, the controller may cause the shaker plate 206 to shake at a second predetermined speed while the vial rack 200 is coupled to the shaker plate 206 and includes at least one vial 208 containing a solution in a vial receptacle 202. Additionally, during this second agitating event, the end of the pipette tip may be positioned at a second predetermined depth within the vial 208. In some examples, the first predetermined depth is the same as the second predetermined depth. Alternately, in some examples, this second predetermined depth is different from the first predetermined depth. Namely, each predetermined depth corresponds to the volume of solution left in the vial 208. Since there will be less solution of in vial 208 after each corresponding agitation and withdrawing event, successive predetermined depths are positioned lower within the vial 208.

This process may be repeated for subsequent withdrawing events. Namely, the controller may cause the shaker plate 206 to agitate the vials 208 and the pipettes 210 to withdraw a volume of solution repeatedly. Each agitation and/or withdrawing in a series may be performed at the same mixing speed, mixing pattern, the end of the pipette tip depth, and/or withdraw the same volume of solution. Alternatively, these various settings may change between each agitation and/or withdrawing event in a series. In some examples, agitation can involve aspirating the solution. In other examples, agitation can involve rapidly withdrawing and dispensing solution.

Additionally, in some example embodiments, the one or more components of the shaker plate system may provide feedback to a user/operator. For example, the plate shaker system may provide an alert signal (e.g., via a user interface of the controller and/or the shaker plate) to provide an indication to the user that the mixing speeds are outside of a predetermined range (e.g., if the mixing speed of the shaker plate 206 is below about 900 rpm or above about 1100 rpm).

In some example embodiments, the controller may cause the shaker plate system to undergo a read buffer phase. Namely, the controller can cause the shaker plate 206 to shake a dispensing plate at a predetermined mixing speed (e.g., 500 rpm) while the withdrawn solution is dispensed. The shaker plate 206 can then be stopped and the dispensing plate with the withdrawn solution can be transported to a reader to conduct the assay. As explained in further detail with respect to FIGS. 5C-5D, the read buffer phase can further disperse the particles before conducting the assay to provide a more accurate particle count.

Now referring to FIGS. 2D-2G, a vial rack 200 according to an example embodiment. The vial rack 200 includes the base 204 coupled to the one or more vial receptacles 202, suitable for holding vials. As described above, the vial receptacles 202 are configured to hold a variety of vials, some, or all of which may meet SBS standards. Similarly, in FIGS. 2D-2G, the vial receptacles 202 are position the vials 208 axially align with the pipettes 210 such that the pipettes 210 can be inserted and take withdrawals from each of the vials 208.

In some example embodiments, the vial receptacles 202 are designed to substantially surround standard SBS vials (i.e., circular shapes), as shown in FIGS. 2A-2D. Alternatively, in another example, the vial receptacles 202 may be designed to partially surround standard SBS vials. In other examples, the vial receptacles 202 may hold standard SBS vials in any suitable manner.

In practice, the vial receptacles 202 work to stabilize vials during agitation and withdrawing of the particle solution. Stabilization of the vials 208 is desirable during withdrawing and agitation of a particle solution to reduce variability among operators by reducing inconsistencies between particle counts over a series of withdrawing events. In some examples, these particles may be microbeads, microparticles, micropellets, microwafers, paramagnetic beads, microparticles, paramagnetic microparticles, or the like. In embodiments, the particles contain one or more identifying features such as a bar code, a nickel bar code, an alphanumeric symbol, a color, or some combination of thereof, among other possibilities. Particle sizes may range from about 70×25×6 micrometers (μm) to about 80×30×6 μm and must be able to be withdrawn into the pipette tip. In embodiments, the vial receptacles 202 are positioned to axially align the vials 208 with the pipettes 210 prior to the agitation and withdrawing events.

In some examples, the base 204 of the vial rack 200 is configured to detachably associate from a SBS shaker plate, such as the Hamilton® Genomic STARlet. For example, the dimensions of the base can be the same or substantially similar to those of the dimensions of the shaker plate.

In some examples, the vial rack 200 includes fastening mechanisms to detachably associate from a shaker plate. In some examples, the fastening mechanisms can include screws and/or a threaded connection compatible with the shaker plate to keep the vial rack 200 in place during agitation and withdrawing. In this regard, the vial rack 200 may be permanently or detachably associated with the shaker plate. Namely, in practice, the vial rack 200 is configured to be permanently or removably mounted to the shaker plate in order to support a pipette in a manner that consistently and repeatedly positions the vial rack 200 (and vials therein) in relation to the pipette. In this manner, the withdrawing events associated with the vial rack, vials, shaker plate, and pipettes may be consistently and repeatedly performed, thereby improving the results of assays and other testing results associated therewith. In a further aspect, the vial rack 200 has multiple configurations to position an end of a pipette tip at a series of predetermined depths within a vial in the vial receptacles.

In some examples, the vial rack 200 includes the one or more vial receptacles 202. In the example shown in FIGS. 2A-2D, the vial rack 200 includes 32 vial receptacles 202, or four rows of eight vial receptacles 202. In practice, this alignment of vial receptacles 202 is desirable because the Hamilton® Genomic STARlet includes 8 pipettes. Accordingly, the four rows of eight vials receptacles 202 allows for conducting four assays of particle solution within the eight vials without requiring stoppage of the machine.

Now referring to FIGS. 3A-3B, a pipette 210 and a vial 208 according to example embodiments and configurations. Namely, FIG. 3A illustrates an end of the pipette tip 212 at a first predetermined depth 302 within the vial 208, with the end of the pipette tip at or near the center or middle of the horizontal cross section of the vial (the “horizontal center”). FIG. 3B illustrates the end of the pipette tip 212 at a second predetermined depth 310 within the vial 208.

In practice, the vial 208 contains a solution 304, which includes particles 306. These particles may be utilized during one or more assay procedures after the solution 304 is withdrawn from the vial 208, including, for example, to identify a particular type and/or subset of particles within the solution.

The first predetermined depth 302 is determined based on the volume of the solution 304 within the vial 208 at or before the first withdrawing event. As noted above, it may be desirable for the end of the pipette tip 212 to be at or near the vertical and/or horizontal center of the solution 304. The vertical center of the solution can be determined based on the size of the vial 208, the volume of solution 304, and/or the first height 308 of the solution 304 during or before the first withdrawing event.

The second predetermined depth 310 is determined based on the volume of the solution 304 within the vial 208 at or before a second withdrawing event. The vertical center of the solution can be determined based on the size of the vial 208, the volume of solution 304, and/or the second height 312 of the solution 304 during or before the second withdrawing event. In practice, the vertical center of the solution 304 during or before the second withdrawing event will be lower than before the first withdrawing event based on how much solution 304 is withdrawn during the first withdrawing event. Similarly, the first height 308 of the solution 304 is higher than the second height 312 of the solution 304 and the second predetermined depth 310 is typically deeper within the vial 208 than the first predetermined depth 302.

As described above, a controller is configured to position the end of the pipette tip 212 at the first predetermined depth 302 during a first withdrawing event. The controller is configured to position the end of the pipette tip 212 at the second predetermined depth 310 during a second withdrawing event, different from the first predetermined depth 302. Although two predetermined depths are shown in FIGS. 3A-3B, it should be understood that many other predetermined depths can be achieved based on various configurations and examples. It should also be noted that although round particles are illustrated in FIGS. 3A-3B, different shapes, amounts, and/or types of particles may be used.

FIGS. 3C-3D show the example embodiment of FIGS. 3A-3B, except the solution 304 includes paramagnetic beads 314. In some example embodiments, each of the paramagnetic beads 314 includes a unique bar code. In another example, each of the paramagnetic beads 314 includes two or more unique bar codes.

In yet another example, a subset of the paramagnetic beads 314 may include one unique bar code and the remaining paramagnetic beads 314 may include two or more unique bar codes. In practice, each of these bar codes may correspond to particular information about the paramagnetic bead, the solution, and/or one or more additional parameters (including those used in an assay). For example, these unique bar codes may be utilized during one or more assay procedures after the solution 304 is withdrawn from the vial 208, including, for example, to identify a particular type and/or subset of paramagnetic beads within the solution.

Like FIGS. 3A-3B, although two predetermined depths are shown in FIGS. 3C-3D, it should be understood that many other predetermined depths can be achieved based on various configurations of the pipette 210, shaker plate 206, and/or vial rack 200.

In a further aspect, to evaluate the efficacy of preparing, homogenizing, and/or counting the particles in a solution containing a plurality of particles one or more of the example embodiments described above, one or more devices, systems, or methods may be employed.

For example, if particles in a solution are not even distributed throughout the solution prior to a withdrawing event (e.g. because of clumping, binding/settling on the bottom or sides of the vessel in which the solution containing the particles is prepared, etc.), the particles may remain in the vessel as solution is pipetted out of the vessel. To help measure the dispersion and consistency of particles in a solution, a sample of the solution may be transferred onto a surface (e.g., a Petri dish, a well, or the like) and a composite image of the transferred sample of solution may be generated. In examples, this composite image may contain a plurality of images of the transferred sample of solution and based on one or more attributes of this generated composite image, one or more parameters may be determined for the transferred sample of solution.

In FIGS. 4A-4D, a sample of solution 402 containing particles is illustrated according to an example embodiment. These particles may be utilized during one or more assay procedures, including, for example, to identify a particular type and/or components within a sample. In the example embodiment illustrated in FIGS. 4A-4D, the solution 402 includes paramagnetic beads 404. In some example embodiments, each of the paramagnetic beads 404 includes a unique identifier, such as a bar code. In another example, each of the paramagnetic beads 404 includes two or more unique identifiers, such as bar codes.

In yet another example, a subset of the paramagnetic beads 404 may include one unique bar code and the remaining paramagnetic beads 404 may include two or more unique bar codes. In practice, each of these bar codes may correspond to particular information about the paramagnetic bead, the solution, and/or one or more additional parameters (including those used in an assay). For example, these unique bar codes may be utilized during one or more assay procedures to identify a particular type and/or subset of paramagnetic beads within the solution.

It should also be noted that although the particles illustrated in FIGS. 4A-4D involve paramagnetic beads, different shapes, amounts, and/or types of particles may be used.

It should also be noted that one or more concepts illustrated in FIGS. 4A-4D may be accomplished using a computing device, such as computing device 100.

In FIG. 4A, a sample of the prepared solution transferred onto a surface is illustrated, wherein the prepared solution comprises the first volume of buffer solution and the plurality of particles. Turning to FIG. 4B, an example segmentation 406 of the surface is illustrated, in which the surface has been divided into four quadrants for imaging. In example embodiments, segmentation of the illustrated surface may include different sizes, shapes, numbers, and configurations of segments to be imaged for the transferred solution, depending on one or more characteristics of the sample (e.g., size, particle concentration, etc.) and/or the image analysis to be undertaken. Once the segmentation 406, one or more images may be captured for each of the one or more segments and used for further processing.

Turning to FIG. 4C, an example composite image 410 of a plurality of images captured across one or more segments of the solution is illustrated. In example embodiments, composite image 410 may be generated by stitching the plurality of images of the transferred sample of prepared solution into the composite image of the transferred sample of solution illustrated in FIG. 4C. In example embodiments, a controller may stich together the plurality of images of the transferred sample of solution that contain particles and remove any images that do not contain particles. In example embodiments, the controller may determine which images in the plurality of images of the transferred sample of solution that contain particles by performing one or more of a pixel density and/or gradient analysis of the individual images captured by the controller. In some example embodiments, the particles in the solution (e.g., paramagnetic beads 404) may present a different contrast and/or pixel density compared the solution in which the particles are disposed (shown in FIGS. 4A-4D as the dark, black portions of paramagnetic beads 404 compared to the light, white portions of the surround solution 402). Prior to stitching, as illustrated in the example graphical user interface 408 of FIG. 4C, a user may set one or more parameters for the stitching protocol, including which images should be stitched together, as well as one or more attributes of the stitched image and/or stitching protocol (shown in FIG. 4C as “Max Corners,” “Min Distance,” “Block Size,” “Block Gradient,” “K”). Other examples are possible.

Once composite image 410 has been generated, further analysis may be undertaken on the composite image 410 to determine one or more parameters of the transferred sample of prepared solution and/or the particles contained therein. In example embodiments, as shown in the example graphical user interface 408 of FIG. 4C, a user may want to determine one or more attributes of the solution, including a count of the particles in the transferred sample. To do so, the user may select to use one or more programs executing a variety of automated protocols, including one or more edge detection protocols. In an example embodiment, as illustrated in FIG. 4C, a user may select to use a Harris corner detection algorithm (shown in FIG. 4C as “Use Harris Detector” prompt 412) to perform this edge detection for the particles in the solution and thereby generate a count of the particles in the solution. In example embodiments, the controller may use one or more algorithms (including Harris corner detection) and/or protocols to detect an edge of a particle in the composite image, based at least in part on detecting an edge of the particle in the composite image, determining a presence of at least one particle in the composite image. Other examples, including the use of other image processing and/or machine learning and artificial intelligence algorithms, are possible. For example, one or more machine learning models may comprise a deep learning model and/or image pixel and/or gradient analysis models.

Turning to FIG. 4D, an example annotated image 414 of a plurality of particles detected in the composite image 410 is illustrated. In example embodiments, the controller may determine the count of the particles in solution by identifying the particles in the solution and generate annotated image 414 by utilizing one or more edge detection protocols (including the Harris corner detection algorithm). In example embodiments, the controller may perform this edge detection for the particles in the solution, generate a count of the particles in the solution, and present annotated images of the particles identified and/or counted in the annotated image 414. For example, as shown in example graphical user interface 416 of FIG. 4D, in example embodiments, the controller may present the user with an image accounting 418 of all of the images that were stitched together and/or annotated, as well as one or more attributes of the stitched image and/or stitching protocol (e.g., how many particles were identified in each image and/or segment of sample and/or corresponding images). Other examples are possible.

Once annotated image 414 has been generated, further actions may be undertaken on the annotated image 414 to further inform a user of the controller of one or more parameters of the transferred aliquot of prepared solution and/or the particles contained therein. In example embodiments, as shown in the example graphical user interface of FIG. 4D, a user may be presented with an annotated version of a single particle 420 in the solution, as well an annotated version of a multiple particles 422 that may be overlapping and/or joined together in the solution. In example embodiments, the user may select to generate one or more additional annotated images and/or graphical user interfaces based on the annotated image 414, including total particle counts in the sample, the types of particles in the sample, and the extent of overlapping particles in the sample.

These example graphical user interfaces are merely for purposes of illustration. The features described herein may involve graphical user interfaces that are configured or formatted differently, include more or less information and/or additional or fewer instructions, include different types of information and/or instructions, and relate to one another in different ways.

EXAMPLES

To illustrate the example embodiments described above, several sample solutions were prepared and tested to measure the efficacy of the mixing speed and pipette depth of the shaker plate system. In example embodiments, these solutions may utilize a proprietary solution, a Phosphate Buffered Saline with Tween solution (PBS-T), some combination of the two, or other solutions. Additionally or alternatively, these solutions may also include a mixture of particles (e.g., bar coded paramagnetic beads) with a variety of different attributes (e.g., all bar coded paramagnetic beads may comprise a single bar code or a mix bar codes). Additionally, several solutions were prepared and tested to measure the efficacy of performing a read buffer phase. Further details are provided below.

Example 1: Mixing Speed

In an example experiment, a Hamilton® Genomic STARlet shaker system was used to aspiration and withdraw a paramagnetic bead solution at different mixing speeds. Namely, a paramagnetic bead solution was withdrawn from a set of vials twelve times at a particular mixing speed. After each of the twelve withdrawals at the particular mixing speed, an assay was conducted to determine paramagnetic bead count. This series of withdrawals was tested with mixing speeds ranging from 600 revolutions per minute (rpm) to 1200 rpm.

FIG. 5A shows the results of this experiment. Namely, FIG. 5A shows the paramagnetic bead count assay results over five different mixing speeds: 600 rpm, 800 rpm, 900 rpm, 1000 rpm, and 1200 rpm.

As shown in FIG. 5A, mixing at 600 rpm and 800 rpm produced a low and/or inconsistent paramagnetic bead count. Mixing at 900 rpm and 1000 rpm produced a higher and more consistent paramagnetic bead count. Mixing at 1200 rpm produced a higher, but less consistent paramagnetic bead count. Mixing at 1200 rpm also caused the solution to produce bubbles which may cause issues with the assay. As shown in FIG. 5A, mixing at between 900 rpm and 1000 rpm produced the most homogenous dispersion of beads throughout the solution and thereby the most consistent bead counts across all vials and all withdrawing events over the vials.

Additionally, utilizing the shaker system to mix the bead solution not only improved bead count and solution homogeneity and consistency across all vials and withdrawing events, it also saves time in the overall assay procedure, which allows the user to increase throughput (whether in an automated or manual procedure) resulting in time and cost savings for the assay procedure.

Furthermore, although the experiment in Example 1 describes particular components and tests utilized according to specific parameters, it should be understood that the claimed devices and/or methods may be implemented in a variety of scenarios, including scenarios other than the assays described herein. For example, claimed devices and/or methods may be implemented in any situation where there is a need to pipette a consistently homogenous solution.

Example 2: Shaking Speed

In an example experiment, a Hamilton® Genomic STARlet shaker system was used to aspiration and withdraw a paramagnetic bead solution at a mixing speed of 1000 rpm. Namely, a paramagnetic bead solution was withdrawn from a set of vials twelve times at 1000 rpm mixing speed. After each of the twelve withdrawals at the particular mixing speed, an assay was conducted to determine paramagnetic bead count.

The pipette tip depth was 0.15 millimeters (mm) from the bottom of the vial during withdrawing events 1-11. The pipette tip depth was 0.12 mm from the bottom of the vial during the 12^(th) withdrawing event. The aspiration volume was 100 microliters (μL) during withdrawing events 1-11. The aspiration volume was 115 μL during the twelfth withdrawing event. The dispense volume was 100 μL during withdrawing events 1-11. The dispense volume was 115 during the twelfth withdrawing event. In this regard, by adjusting both the volume and pipette depth in the twelfth withdrawing event, bead count consistency was improved as compared to the other eleven withdrawing events, thereby resulting in an improved overall assay. The mix speed was held constant at 1000 rpm for each of the twelve withdrawing events.

As shown in FIG. 5B, the mixing conditions described above produced consistent paramagnetic bead counts near the target of 400 paramagnetic beads per well.

Furthermore, although the experiment in Example 2 describes particular components and tests utilized according to specific parameters, it should be understood that the claimed devices and/or methods may be implemented in a variety of scenarios, including scenarios other than the assays described herein. For example, claimed devices and/or methods may be implemented in any situation where there is a need to pipette a consistently homogenous solution.

Example 3: Read Buffer Phase

In an example experiment, paramagnetic bead solution was withdrawn and dispensed into a stationary dispense plate and then a dispense plate positioned on a Hamilton® Genomic STARlet shaker plate at a mix speed of 500 rpm. In both instances, 200 μL of read buffer solution was dispensed at a dispense rate of 300 μL per second (i.e., the total volume of the 200 μL of solution was dispensed into the vial in 0.666 seconds).

FIG. 5C shows the paramagnetic bead dispersion after the 200 μL of read buffer solution was dispensed in the vials on the stationary plate. FIG. 5D shows the paramagnetic bead dispersion after the 200 μL of read buffer solution was dispensed in the vials on the dispensing plate at a mixing speed of 500 rpm. By shaking the plate at 500 rpm, the read buffer solution was able to dispense into all locations of the interior well of the vials on the dispense plate and disperse the randomly placed paramagnetic beads in the total solution. In this regard, the total bead dispersion improved in the final solution, as did the results of the assay based on increased bead count. For example, as shown in FIG. 5C, with the stationary plate, 50% of the paramagnetic beads were counted in the assay. For comparison, in FIG. 5D, with the dispensing plate at a mixing speed of 500 rpm, 80% of the paramagnetic beads were counted in the assay.

Furthermore, although the experiment in Example 3 describes particular components and tests utilized according to specific parameters, it should be understood that the claimed devices and/or methods may be implemented in a variety of scenarios, including scenarios other than the assays described herein. For example, claimed devices and/or methods may be implemented in any situation where there is a need to pipette a consistently homogenous solution.

Example 4: Automated Bead Count

In an example experiment, using the devices, systems, or methods of FIGS. 4A-4D, a controller and associated program instructions to determine the accuracy of an automated bead counting protocol and compare it to a manual bead counting protocol. In the first portion of the experiment, an automated bead counting protocol was used to capture, stich, and analyze images of the sample solution to determine the total number (counts) of particles in the solution sample contained in a well (a Petri dish). In a second portion of the example experiment, the sample solution was analyzed manually by an operator to determine the total number (counts) of particles in the solution sample contained in a well (a Petri dish), by manually counting the particles in the captured images. After the first and second portions of the experiment, the counts were compared to determine an agreement between the automated and manual bead counting protocols.

FIG. 5E shows the results of this experiment. Namely, FIG. 5E shows 99% agreement between the automated and manual bead counting protocols.

Additionally, utilizing automated bead counting protocols improved bead count consistency across image analysis, it also saves time in the overall bead counting protocol and solution preparation procedure, as well as ensures more consistent results in the solution preparation procedure. For example, because a user may efficiently and consistently test one or more parameters of the prepared solution (e.g., bead counts in representative samples of the prepared solution), the may user to increase the throughput (whether in an automated or manual procedure) and consistency of prepared solution, resulting in time and cost savings, as well as improved results, for the solution preparation (and assay) procedure.

Furthermore, although the experiment in Example 4 describes particular components and tests utilized according to specific parameters, it should be understood that the claimed devices and/or methods may be implemented in a variety of scenarios, including scenarios other than the solution preparation and/or assays described herein. For example, claimed devices and/or methods may be implemented in any situation where there is a need to prepare and/or pipette a consistently homogenous solution.

Example Methods and Aspects

Now referring to FIG. 6 , an example method of withdrawing a solution including a plurality of particles is illustrated. Method 600 shown in FIG. 6 presents an example of a method that could be used with the computing device 100, vial rack 200, shaker plate 206, and pipette 210 with a pipette tip 212 shown in FIGS. 1-5E, for example. Further, devices or systems may be used or configured to perform logical functions presented in FIG. 6 . In other examples, components of the devices and/or systems may be arranged to be adapted to, capable of, or suited for performing the functions, such as when operated in a specific manner. Method 600 may include one or more operations, functions, or actions as illustrated by one or more of blocks 602-604. Although the blocks are illustrated in a sequential order, these blocks may also be performed in parallel, and/or in a different order than those described herein. Also, the various blocks may be combined into fewer blocks, divided into additional blocks, and/or removed based upon the desired implementation.

At block 602, method 600 for withdrawing a solution comprising a plurality of particles from a vial involves agitating the vial at a first predetermined mixing speed to suspend the plurality of particles in the solution.

In some example embodiments, agitating the vial at the first predetermined mixing speed comprises agitating the vial in an orbital pattern. Alternatively, agitating the vial at the first predetermined mixing speed comprises agitating the vial in a linear pattern. Further, in some examples, the first predetermined mixing speed is between 900 revolutions per minute and 1100 revolutions per minute.

At block 604, method 600 involves withdrawing, during the agitating, a first volume of the solution from the vial with a pipette, such that an end of a pipette tip is positioned at a first predetermined depth within the vial.

In examples, the plurality of particles comprise a plurality of paramagnetic beads. Additionally, in some examples, the plurality of paramagnetic beads comprise two or more unique bar codes.

In some example embodiments, method 600 involves, prior to withdrawing and during the agitating, the first volume of the solution from the vial, aligning the pipette with the vial along an axis using a vial rack, wherein the vial rack is configured to receive and stabilize the vial.

Additionally, in some examples, the method 600 further includes agitating the vial at a second predetermined mixing speed to suspend the plurality of particles in the solution. In these examples, the method 600 can additionally include withdrawing, during the agitating, a second volume of the solution from the vial with the pipette, wherein the pipette tip is positioned at a second predetermined depth within the vial, and wherein the second predetermined depth is different than the first predetermined depth as result of a volume of the solution remaining in the vial. In these examples, the second predetermined depth is determined based on measuring a distance from the pipette tip to a bottom portion of the vial.

Further, in some examples, the first predetermined mixing speed and the second predetermined mixing speed are the same. Alternatively, in some examples, the first predetermined mixing speed and the second predetermined mixing speed are different.

Additionally, in some examples, the first volume and the second volume may be the same. Alternatively, in some examples, the first and second volumes are different.

In one aspect, a non-transitory computer-readable medium, having stored thereon program instructions that, upon execution by a controller, cause a controller to perform a set of operations comprising agitating a vial at a first predetermined mixing speed to suspend a plurality of particles in a solution and withdrawing, during the agitating, a first volume of the solution from a vial with a pipette, such that an end of a pipette tip is positioned at a first predetermined depth within a vial, is disclosed.

In example embodiments, the set of operations further comprises agitating a vial at a second predetermined mixing speed to suspend the plurality of particles in the solution, and withdrawing, during the agitating, a second volume of the solution from a vial with a pipette, wherein a pipette tip is positioned at a second predetermined depth within a vial, and wherein the second predetermined depth is different than the first predetermined depth as result of a volume of the solution remaining in a vial, and wherein the second predetermined depth is determined based on measuring a distance from a pipette tip to a bottom portion of a vial.

In example embodiments, the set of operations further comprises determining that a vial is agitated outside of a predetermined range of the first predetermined mixing speed or second predetermined mixing speed, and generating an alert signal to provide an indication that a vial is agitated outside of the predetermined range of the first predetermined mixing speed or second predetermined mixing speed. In example embodiments, the set of operations further comprises, based on determining that a vial is agitated outside of a predetermined range of the first predetermined mixing speed or second predetermined mixing speed, stopping agitation of a vial.

In example embodiments, the plurality of particles comprises a plurality of paramagnetic beads, wherein the plurality of paramagnetic beads comprises one or more unique bar codes.

In some example embodiments, agitating a vial at the first predetermined mixing speed comprises agitating a vial in an orbital pattern, and wherein the first predetermined mixing speed is between 900 revolutions per minute and 1100 revolutions per minute. In example embodiments, prior to withdrawing, during the agitating, the first volume of the solution from a vial, aligning a pipette with a vial along an axis using a vial rack, wherein a vial rack also stabilizes the vial.

The singular forms of the articles “a,” “an,” and “the” include plural references unless the context clearly indicates otherwise. For example, the term “a compound” or “at least one compound” can include a plurality of compounds, including mixtures thereof.

Various aspects and embodiments have been disclosed herein, but other aspects and embodiments will certainly be apparent to those skilled in the art. Additionally, the various aspects and embodiments disclosed herein are provided for explanatory purposes and are not intended to be limiting, with the true scope being indicated by the following claims. 

What is claimed is:
 1. A method for withdrawing a solution comprising a plurality of particles from a vial, the method comprising: agitating the vial at a first predetermined mixing speed to suspend the plurality of particles in the solution; and withdrawing, during the agitating, a first volume of the solution from the vial with a pipette, such that an end of a pipette tip is positioned at a first predetermined depth within the vial.
 2. The method of claim 1, wherein the plurality of particles comprises a plurality of paramagnetic beads.
 3. The method of claim 2, wherein the plurality of paramagnetic beads comprises one or more unique bar codes.
 4. The method of claim 1, wherein agitating the vial at the first predetermined mixing speed comprises agitating the vial in an orbital pattern.
 5. The method of claim 4, the method further comprising, prior to the agitating, inserting the end of the pipette tip at or near the center of a horizontal cross section of the vial, and wherein during the agitating, the orbital pattern comprises a pattern of up to 3 millimeters from horizontal center.
 6. The method of claim 4, wherein the first predetermined mixing speed is between 900 revolutions per minute and 1100 revolutions per minute.
 7. The method of claim 1, wherein agitating the vial at the first predetermined mixing speed comprises agitating the vial in a linear pattern.
 8. The method of claim 1, the method further comprising: prior to withdrawing, during the agitating, the first volume of the solution from the vial, aligning a pipette with the vial along an axis using a vial rack, wherein the vial rack is configured to receive and stabilize the vial.
 9. The method of claim 1, the method further comprising: agitating the vial at a second predetermined mixing speed to suspend the plurality of particles in the solution; and withdrawing, during the agitating, a second volume of the solution from the vial with the pipette, wherein the pipette tip is positioned at a second predetermined depth within the vial, and wherein the second predetermined depth is different than the first predetermined depth as result of a volume of the solution remaining in the vial.
 10. The method of claim 9, further comprising, prior to withdrawing the second volume, the second predetermined depth, wherein the second predetermined depth is determined based on measuring a distance from the pipette tip to a bottom portion of the vial.
 11. The method of claim 9, wherein the first predetermined mixing speed and the second predetermined mixing speed are the same.
 12. The method of claim 9, wherein the first predetermined mixing speed and the second predetermined mixing speed are different.
 13. The method of claim 9, wherein the first volume and the second volume are the same.
 14. The method of claim 9, wherein the first volume and the second volume are different.
 15. The method of claim 1, wherein the method further comprises: transferring a sample of the solution onto a surface; generating a composite image of the transferred sample of solution, wherein the composite image comprises a plurality of images of the transferred sample of solution; and based on the generated composite image, determining a parameter of the transferred sample of solution.
 16. The method of claim 15, wherein generating the composite image of the transferred sample of solution further comprises stitching the plurality of images of the transferred sample of solution into the composite image of the transferred sample of solution.
 17. The method of claim 15, wherein determining a parameter of the transferred sample of solution comprises counting the plurality of particles in the transferred sample of solution.
 18. The method of claim 17, wherein counting a plurality of particles in the transferred sample of solution comprises: detecting an edge of a particle in the composite image; and based at least in part on detecting an edge of the particle in the composite image, determining a presence of at least one particle in the composite image.
 19. A non-transitory computer-readable medium, having stored thereon program instructions that, upon execution by a controller, cause a controller to perform a set of operations comprising: agitating a vial at a first predetermined mixing speed to suspend a plurality of particles in a solution; and withdrawing, during the agitating, a first volume of the solution from the vial with a pipette, such that an end of a pipette tip is positioned at a first predetermined depth within the vial.
 20. The non-transitory computer-readable medium of claim 19, wherein the plurality of particles comprises a plurality of paramagnetic beads, wherein the plurality of paramagnetic beads comprises one or more unique bar codes.
 21. The non-transitory computer-readable medium of claim 19, wherein agitating the vial at the first predetermined mixing speed comprises agitating the vial in an orbital pattern, and wherein the first predetermined mixing speed is between 900 revolutions per minute and 1100 revolutions per minute.
 22. The non-transitory computer-readable medium of claim 19, the set of operations further comprising: prior to withdrawing, during the agitating, the first volume of the solution from the vial, aligning the pipette with the vial along an axis using a vial rack, wherein the vial rack also stabilizes the vial.
 23. The non-transitory computer-readable medium of claim 19, the set of operations further comprising: agitating the vial at a second predetermined mixing speed to suspend the plurality of particles in the solution; and withdrawing, during the agitating, a second volume of the solution from the vial with the pipette, wherein the pipette tip is positioned at a second predetermined depth within the vial, and wherein the second predetermined depth is different than the first predetermined depth as result of a volume of the solution remaining in the vial, and wherein the second predetermined depth is determined based on measuring a distance from the pipette tip to a bottom portion of the vial.
 24. The non-transitory computer-readable medium of claim 23, the set of operations further comprising: determining that the vial is agitated outside of a predetermined range of the first predetermined mixing speed or second predetermined mixing speed; and generating an alert signal to provide an indication that the vial is agitated outside of the predetermined range of the first predetermined mixing speed or second predetermined mixing speed.
 25. The non-transitory computer-readable medium of claim 21, the set of operations further comprising: based on determining that the vial is agitated outside of a predetermined range of the first predetermined mixing speed or second predetermined mixing speed, stopping agitation of the vial. 